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bacterial endotoxin test

bacterial endotoxin test

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Published by ankzii

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Categories:Types, Research
Published by: ankzii on Jun 21, 2009
Copyright:Attribution Non-commercial

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05/24/2013

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BACTERIAL ENDOTOXIN TEST
PRINCIPLE
 
BET measures the concentration of bacterial endotoxin that may be presentin article to be tested. The detection of the Gram negative bacterialendotoxin are based on the use of a Limulus Amoebocytes Lysate (LAL)obtained from the aqueous extract of circulating amoebocyte of horse shoecrab (
Limulus polyphemus) used as LAL Reagent 
 
THEORY 
LAL assay is based on the observation that when an endotoxin contacts theclotable protein from circulating amoebocytes of Limulus, a gel clot forms.The assay kits contain calcium proclotting enzyme and procoagulogen. Theproclotting enzyme is activated by bacterial endotoxin lipopolysaccharideand calcium to form active clotting enzyme. Active clotting then catalyzesthe procoagulogen into polypeptide subunits (coagulogen). The subunit joinsby disulfide bond to form a gel clot .
 
o
ProclottingEnzyme(
inactive
)
Endotoxin SampleClotting Enzyme(
active )
-----------------------||vProcoagulogen(
soluble
)--------------Coagulogen(
insoluble
)|||||vGel clot

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